Prick Test Research Articles

PPARγ alleviates damage to chorionic trophoblast cells induced by high glucose and high lipids through regulation of IGF-1.

Overweight and obesity are the most common high-risk conditions that increase the risk of adverse outcomes during pregnancy, childbirth, and the postpartum period. Dysfunctions in trophoblastic peroxisome proliferator-activated receptor gamma (PPARγ) contribute to a variety of related pregnancy disorders. This study investigated whether PPARγ influences chorionic trophoblast cell damage induced by high glucose (HG) and high lipid (HL) by regulating insulin-like growth factor-1 (IGF-1). Human trophoblast HTR-8/SVneo cells were exposed to HG and HL conditions to simulate damaged trophoblasts during pregnancy in vitro. Cell Counting Kit-8 (CCK-8) was used to assess cell proliferation. The Scratch test was used to test cell migration. Cell invasion ability was assessed by Transwell assay. ELISA was used to assess the inflammatory factor levels. Glucose, lactic acid, and adenosine triphosphate (ATP) levels were measured using biochemical kits. High glucose/HL inhibited the proliferation, migration, and invasion of HTR-8/SVneo cells. High glucose and HL increased tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, and IL-6 expression while decreasing IL-10 expression. High glucose and HL decreased glucose uptake and ATP levels. High glucose and HL reduced the expressiofns of PPARγ, IGF-1, insulin receptor substrate (IRS) 1, IRS2, GLUT1, and GLUT4. High PPARγ expression promoted cell proliferation, migration, and invasion induced by HG and HL, increased glucose uptake and ATP levels and inhibited inflammation. Low IGF-1 expression inhibited cell proliferation, migration, and invasion under HG and HL conditions, reduced glucose uptake and ATP levels, and increased inflammation. Low IGF-1 expression reversed the effects of PPARγ on HTR-8/SVneo cells under HG and HL conditions. Peroxisome proliferator-activated receptor gamma alleviated HTR-8/SVneo cell damage induced by HG and HL by regulating IGF-1, suggesting a potentially effective approach for treating gestational obesity.

Worse lung function, more allergic sensitization but less blood eosinophilia in elderly patients with long-standing versus late-onset asthma

ABSTRACT Background Asthma in the elderly is usually considered homogeneous and non-atopic. Objective To compare clinical, functional and immunological features between elderly asthmatics with long-standing asthma (LSA) and those with late-onset asthma (LOA). Methods Eighty-two asthmatics older than 64 were included into LSA (asthma onset before age 40; n =46) and LOA (asthma onset from 40 years of age on; n = 36) groups. Asthma treatment and comorbidities were recorded. All individuals underwent the asthma control questionnaire-7 (ACQ-7) and cognitive impairment screening (Mini-Mental State Examination). Inhaler technique was assessed by checklists; the Morisky Medication Adherence Scale-8 was used to assess adherence to treatment. Spirometry, skin prick tests (SPTs), induced sputum and blood eosinophil counts were performed. Results We found high frequencies of cognitive impairment, poor inhaler technique and low adherence to treatment in both groups, which had good disease control (ACQ-7 scores: 1.20 ± 0.74 versus 1.11 ± 0.89; p = 0.67, respectively). The LSA group had more severe airway obstruction (FEV1(% predicted): 62.04 ± 19.50 versus 77.15 ± 18.74, p< 0.01; FEV1/FVC: 0.59 ± 0.10 versus 0.69 ± 0.09, p < 0.01); higher frequency of positive SPTs (65.6% versus 18.8%, p = 0.001); and lower frequency of blood eosinophilia (45.7% versus 77.1%, p = 0.004) than the LOA group. No differences in sputum cell counts or inflammatory profiles were found between the groups. Ninety percent of the individuals studied had at least one feature of Type 2 asthma. Conclusion LSA and LOA phenotypes differ substantially. That should be accounted for in research and clinical practice grounds.

Clinical and sensitization profile in peach allergy due to LTP sensitization

BackgroundLipid transfer proteins (LTP) are associated with a wide range of severity of allergic reactions. However, the risk factors associated with this severity are not fully understood.ObjectivesTo describe the clinical characteristics of peach-allergic patients due to LTP sensitization and analyze the relationship between the severity of the reactions and patients’ sensitization profiles.MethodsA retrospective study of peach-allergic patients was performed. Patients were classified into LTP-monoallergic (only peach allergy) or LTP-Syndrome (peach allergy and allergy to other plants-foods related with LTP). Symptoms with Rosaceae family and other related plant foods, skin prick tests (SPTs), and IgE values were recorded.ResultsSeventy-one patients were included, 46.5% suffering from anaphylaxis, 32.3% from urticaria angioedema, and 21.2% from oral allergy syndrome. Six had monoallergy to LTP and 65 LTP syndrome. Clinical severity showed no differences according to peach SPT wheal size or Pru p 3 IgE levels. We also found no differences between the components of LTP-containing foods analyzed, the number of LTPs recognized, and the severity of symptoms. However, anaphylaxis was more frequent in patients with concomitant allergies to ≥3 groups of plant foods.ConclusionsLTP syndrome was the most common presentation in patients with Rosaceae food allergy. The severity of the reactions was not related to peach SPT wheal size or sIgE levels to Pru p 3, but concomitant allergies to ≥3 plant food groups could be a good marker of severity.

Preparation and properties of Ni-SiO2 superhydrophobic coating by brush plating on inner wall of pipeline

Coal is easy to stay on the inner wall of the pipeline during transportation. In order to solve this problem, it is usually necessary to improve the hydrophobicity of the inner wall of the pipeline. In this study, 45 # steel, which is wear-resistant and has good machinability, is used as the matrix for coal pipeline. Ni-SiO2 composite coatings were prepared on the inner surface of 45 # steel pipelines using a novel brush plating tool, and the hydrophobicity of the coatings were evaluated. Firstly, a series of four-factor and three-level orthogonal tests were conducted to investigate the impact of brush plating working voltage (12V, 14V, 16V), nanoparticle concentration (1g/L, 2g/L, 3g/L), brush plating duration (2min, 3.5min, 5min), and bath temperature (25 ℃, 35 ℃, 45 ℃) on the hydrophobicity of the composite coating. Subsequently, optimum process parameters were determined. Then 1H, 1H, 2H, 2H- perfluorodecyltrimethoxysilane was used to modify the coating surface in order to further improve its hydrophobicity. Scanning electron microscope (SEM), energy spectrum analyzer (EDS), optical contact angle measuring instrument, white light interferometer, Vickers hardness tester, scratch tester, friction and wear meter and electrochemical workstation were used to evaluate the coating's related properties. The experimental results show that the Ni-SiO2 nano-composite coating has a higher angle (135°) when the operational voltage of brush plating is 14V, the amount of nanoparticles is 3g/L, the brush plating time is 2min, and the bath temperature is 35 ℃. The concentration of nanoparticles is the biggest factor affecting the hydrophobicity through orthogonal test analysis, so the control variable method is used to further study the influence of the concentration of nanoparticles on the comprehensive performance of the coating. It was found that with the increase of nanoparticle concentration, surface quality and comprehensive performance of the coating showed a trend of increasing first and then decreasing, and the surface of the coating was rougher when the nanoparticles were added at 4g/L, and the maximum porosity was 31.34%. At this time, the contact angle of the super hydrophobic coating has reached 150.1°, which has reached the super hydrophobicity, and the coating has good hardness, wear resistance and corrosion resistance.

Biodegradation and mechanical performance of Silane-chitosan-graphene oxide composite coating on AZ31 magnesium alloys for biomedical applications.

Biodegradable magnesium alloy medical implants have attracted considerable interest thanks to their remarkable biocompatibility and mechanical properties. However, the rapid corrosion rate of magnesium alloys in physiological environments presents a major challenge to their practical application. Therefore, this study attempted to design a silane/chitosan /graphene oxide composite coating that reduces the corrosion and enhances the biodegradation of magnesium alloys used in temporary implants. The composite fabrication process adopted a cost-effective spin-coating technique providing a uniform coating of magnesium alloy substrates. Fourier transform infrared spectroscopy (ATR-FTIR), Raman spectroscopy, scanning electron microscopy (SEM) and energy dispersion spectroscopy (EDS) results showed a dense and compact silane/chitosan/graphene oxide (Silane/CS/GO) coating on the magnesium coated alloy surface. The mechanical properties of the fabricated silane/chitosan/graphene oxide composite were also investigated by microindentation and scratch tests. The hardness of the magnesium alloy increased from ~268 HV ±13.4 to ~644.13 HV ± 32.2 upon the application of the coating. Moreover, the results showed that the cohesive critical load (LC) of the ternary composite was greater than LC ~ 3.02 ± 0.15 N. In addition, compared to uncoated AZ31 alloys, the friction coefficient and wear volume of the ternary composite decreased from ~0.13 ± 0.02 to ~0.07 ± 0.004 and from ~18 to ~0.04 (106μm3), respectively. Finally, the corrosion of the composite coating was assessed in simulated body fluid (SBF) in vitro at 37 °C. Hydrogen evolution results revealed that graphene oxide seemed to delay the diffusion of corrosive ions into the Mg matrix, while Silane prevented the coupling of GO graphene oxide sheets to the metal surface. Consequently, silane / chitosan /graphene oxide composite coatings could be a very useful material for coating magnesium implant devices. It contributed to reduce the corrosion of the substrate and alleviate the cost and discomfort of implants.

A comparative evaluation of green surfactant-assisted calcium phosphate (CaP) coatings developed through electrodeposition and biomimetic routes

This study compares the properties of green surfactant-assisted calcium phosphate (CaP) coatings developed on 316L stainless steel (316L SS) substrates using electrodeposition (ED) and biomimetic (BM) methods. The influence of biosurfactants (BS) on these coatings' properties, deposited through both processes, is also investigated. X-ray diffraction (XRD), scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS), and Fourier-transform infrared spectroscopy (FTIR) confirm the presence of brushite and hydroxyapatite (HAp) in the ED coatings, while BM coatings show only HAp without BS and both brushite and HAp phases with BS. The SEM analysis reveals distinct morphologies, with BS-assisted coatings showing more organized structures. The presence of BS results in more uniform coatings in both ED and BM methods. Atomic force microscopy (AFM) analysis indicates that BM coatings are smoother than ED coatings, and BS further reduces surface roughness. The comprehensive evaluations through nanoindentation, scratch testing, and corrosion studies reveal that BS-assisted ED coatings exhibit superior hardness (H), elastic modulus (E), adhesion strength, and corrosion resistance. Cytocompatibility studies using the MTT assay demonstrate that all coatings support cell attachment and proliferation without cytotoxic effects, with BM coatings showing slightly better cell viability. This study highlights the potential of green surfactant-assisted CaP coatings developed using ED and BM methods, for improving the biocompatibility and other biological properties of 316L SS implants.

122 Evaluation of the relevance of skin prick test, intradermal test, specific IgE and total IgE levels considering hymenoptera allergy using an own clinical classification

122 Evaluation of the relevance of skin prick test, intradermal test, specific IgE and total IgE levels considering hymenoptera allergy using an own clinical classification

High-temperature abrasive wear behaviour of strengthened iron-aluminide laser claddings

Strengthened iron-aluminides (Fe3Al-based) show notable mechanical properties alongside good scratch hardness up to 600 °C. To investigate the influence of strengthening mechanisms for alternative high temperature wear resistant materials, laser metal deposited alloys based on Fe with 30 at.% Al were studied in detail. After cladding the Fe30Al base material and its alloys – with either 5 at.% Si, 5 at.% C or 3 at.% Ti combined with 6 at.% B – thorough investigations by scanning electron microscopy, electron backscatter diffraction, nanoindentation, hot hardness measurements, and high temperature scratch tests were performed.All alloyed Fe3Al-based claddings show increased strength. The highest overall hardness of ~405 HV10 exhibits the carbon-alloyed one containing a high fraction of Fe3AlC0.6 carbides. Contrary, the Ti + B alloyed as well as the Si-alloyed claddings are superior with their scratch hardness (especially at elevated temperatures), through the formation of TiB2 precipitates and pronounced solid solution strengthening, respectively. Wear tests reveal decreasing wear rates with increasing test temperatures (from 20 to 500 to 700 °C) for all alloyed Fe3Al-based claddings due to the formation of abrasive-containing mechanically mixed layers. Contrary, the reference cladding, cobalt-based Stellite 21 exhibits not only a higher wear rate but also increased ones upon increasing the test temperature. Detailed investigations show that the scratch hardness correlates with the wear resistance and that the incorporation of abrasive material into the wear-induced formation of mechanically mixed layers is highly beneficial. Due to this self-protection effect, the promising overall high temperature behaviour, combined with the alloying concept (no Co, Ni, or Cr), these alloyed Fe3Al-based claddings are promising candidates for sustainable wear protection outperforming currently used wear protection solutions.

Correlation between the structural, morphological, optical, nanomechanical, and tribological properties of ZnO films by USPT with their photocatalytic activities

The present work focused on studying the nanomechanical properties of ZnO thin films deposited on glass substrates using the ultrasonic spray pyrolysis technique. Structural and morphological properties are related to processing conditions, which affect mechanical, optical, and photocatalytic properties. To produce the ZnO films, three different molarities of spraying solution have been used which are 0.05 M; 0.075; and 0.10. The increase in the concentration of the spray solution led to an improvement in the crystal structure with the average crystallite size increasing from 28 nm to 35 nm. Morphological investigations revealed that the spherical nanoparticles with various size distributions. The rough surface topology emerged as a result of increasing solution molarity. Optical band gap values of the films decreased from 3.29 to 3.27 eV. Photocatalytic efficiency reached 93 % for ZnO film produced at higher solution molarity. Improved hardness values were determined for all films. Scratch tests showed better adhesion behavior for ZnO film at low solution molarity. Results indicate that solution molarity is an important parameter of the physical and photocatalytic properties of ZnO films as well as mechanical behaviors.

The Cough Response to Inhaled Mannitol in Healthy Subjects

PurposeInhaled mannitol induces bronchoconstriction and cough. This study aimed to describe the cough response to mannitol among healthy adult subjects.Methods125 healthy subjects (aged 18–82 years, 52% females, 50% skin prick test positive) underwent a mannitol test. The coughs were recorded both simultaneously and afterwards from video recordings by two researchers. Three indices were evaluated: The cumulative number of coughs per cumulative dose of mannitol (CDR), cumulative provocative dose of mannitol to cause at least 5 coughs, and the maximal number of coughs provoked by any single mannitol dose. The test was repeated in 26 subjects after 3–7 days.ResultsCDR showed the best repeatability with an intraclass correlation coefficient of 0.829. Gender was the only characteristics that associated with the cough response: The median CDR was 2.53 (interquartile range 0.45–7.01) coughs/100 mg among females and 0.787 (0.0–3.29) coughs/100 mg among males (p = 0.002). The interquartile range upper limits were defined as the cut-off limits for a normal response. The threshold for a statistically significant change in CDR was 6.26 coughs/100 mg. There was a close correlation between simultaneous- and video-assessed CDR (intraclass correlation coefficient 0.985).ConclusionFemales cough more than males in response to mannitol. CDR is the most suitable index to describe the cough responsiveness. The repeatability of the response is good. Video recording of the coughs is not mandatory. The cut-off limits for a normal cough response to mannitol were provided.

LuLIPLEX: A Fast, Highly Sensitive, and Multiplexed Method for the Detection of IgE Against Major Allergens.

Diagnosis of allergies is mostly based on the patient's clinical history and allergen provocation tests. Determination of specific IgE (sIgE) profiles can be performed to support allergy diagnosis. This is commonly done invivo by the skin prick test or invitro with automated systems. Several platforms exist to quantify sIgE levels, but all these methods require access to specific instruments, often delaying the test's results. The IgE luciferase-linked immunosorbent assay (LuLISA) allows bioluminescent quantification of IgE against peanut in microliter samples, but this method awaits extension to other allergens. This study aimed to validate a new method, named LuLIPLEX, for multiplexed bioluminescent detection of sIgE against 20 major molecular allergens. Quantification of sIgE against 12 recombinant or purified food allergens and eight aeroallergens was performed by LuLIPLEX versus standard IgE detection methods (ImmunoCAP, ISAC, ALEX, or NOVEOS). Multiplexed detection of IgE against these 20 allergens was performed within 45 min using 50 μL of serum, plasma, or whole blood samples. A head-to-head comparison between LuLIPLEX and standard IgE detection methods showed a high correlation among all allergens tested. sIgE profiles in polysensitized subjects could be determined within 45 min in serum and plasma samples, as well as using a single drop of capillary blood. LuLIPLEX is a rapid and sensitive method to quantify sIgE levels against multiple allergens. Given that the test is very fast and can be performed on small and inexpensive luminometers, the IgE LuLIPLEX could allow point-of-care testing of sIgE profiles in allergic subjects.

Efficacy of Procaine, With and Without Epinephrine, Compared to Lidocaine in Local Anesthesia for Calves Before Thermocautery Disbudding.

Within the European Union, the use of lidocaine in food-producing animals is restricted due to concerns over human safety. This study compared the clinical effectiveness of procaine, with and without epinephrine, against lidocaine in pain alleviation during thermocautery disbudding in xylazine-sedated calves. The efficacy of local blocks was assessed through needle pricks, and the behavioral reactions to disbudding were scored. Post-disbudding pain was subjectively evaluated, and pressure pain threshold and tactile sensitivity around the horn bud were assessed at intervals. Blood was collected at intervals for plasma cortisol analysis. No significant differences were found between the groups in the needle prick test (p = 0.329) and the disbudding score (p = 0.855). Pain scores and quantitative sensory tests showed no significant differences between the lidocaine and procaine-epinephrine groups. Conversely, tactile sensitivity and pain scores were significantly higher, and pressure pain thresholds were significantly lower with procaine alone than in other groups. Elevated cortisol concentrations were observed in all groups before disbudding compared to the baselines. The results suggest that procaine combined with epinephrine appears to be a safe and effective alternative to lidocaine for calf disbudding. Cortisol concentrations as an indicator of pain in xylazine-sedated calves appear inadequate.

Study of Cytotoxicity of Spiro-Fused [3-Azabicyclo[3.1.0]hexane]oxindoles and Cyclopropa[a]pyrrolizidine-oxindoles Against Tumor Cell Lines

Background: A series of spiro-fused heterocyclic compounds containing cyclopropa[a]pyrrolizidine-2,3′-oxindole and 3-spiro[3-azabicyclo[3.1.0]-hexane]oxindole frameworks were synthesized and studied for their in vitro antiproliferative activity against human erythroleukemia (K562), cervical carcinoma (HeLa), acute T cell leukemia (Jurkat), melanoma (Sk-mel-2) and breast cancer (MCF-7) as well as mouse colon carcinoma (CT26) cell lines. Methods: Cell proliferation was evaluated in vitro by MTS assay. Confocal microscopy was used to study actin cytoskeleton structure and cell motility. Cell cycle analysis was evaluated by flow cytometry. Results: It was found that compounds 4, 8, 18 and 24 showed antiproliferative activity against the Jurkat, K-562, HeLa and Sk-mel-2 cell lines with IC50 ranging from 2 to 10 μM (72 h). Evaluation of the impact on cell cycle progression showed that the tested compounds achieved significant cell-cycle perturbation with a higher accumulation of cells in the SubG1 and G0/G1 phases of the cell cycle, in comparison to the negative control. I Incubation with tested compounds led to the disappearance of stress fibers (granular actin was distributed diffusely in the cytoplasm in up to 38% of treated HeLa cells) and changes in the number of filopodia-like deformations (reduced from 93% in control cells to 64% after treatment). The impact on the Sk-mel-2 cell actin cytoskeleton structure was even greater: granular actin was distributed diffusely in the cytoplasm in up to 90% of treated cells while the number of filopodia-like deformations was reduced by up to 23%. A scratch test performed on the human melanoma cell line showed that these cells did not fill the scratched strip and lose their ability to move under treatment. Conclusions: The obtained results support the antitumor effect of the tested spiro-compounds and encourage the extension of this study in order to improve their anticancer activity as well as reduce their toxicological risks.

Revolutionizing copper pipe durability: Shielding against corrosion with graphene oxide through electrophoretic application

This research endeavors to utilize electrophoretic deposition (EPD) to coat copper (Cu) pipes with graphene oxide (GO) nanosheets, aiming to bolster their resistance against corrosion. To achieve this, a stable aqueous colloidal suspension of GO was meticulously prepared via liquid exfoliation in deionized water. This suspension served as an electrolytic solution for EPD. The EPD process involved the meticulous application of varied operational parameters such as deposition time, applied voltage, and GO nanosheet concentration to facilitate anodic deposition on the copper pipe’s surface. To gauge the efficacy of the GO coating, evaluations were conducted to assess adhesive force and stabilization using specialized coating adhesion scratch testing equipment. Further analysis of the resulting film on the Cu pipe was executed employing X-ray diffraction (XRD), scanning electron microscopy (SEM), Raman spectroscopy, and Fourier transform infrared spectroscopy (FT-IR). These methods provided detailed insights into the characteristics and properties of the GO material post-deposition. The study yielded noteworthy outcomes, revealing the achievement of a uniform, unbroken, and consistent coating film on the Cu pipe. This was accomplished by employing specific parameters as 60 s of deposition time, a GO concentration of 0.5 mg/mL, and a voltage of 20 V. Impressively, when subjected to a corrosive solution containing 3.5% sodium chloride, the corrosion protection exhibited a remarkable twofold enhancement compared to untreated copper tubing, boasting an efficiency (η) of 90.48%. These findings suggest that the Cu pipe, coated with GO using the EPD technique, holds significant promise for utilization in demanding industrial settings susceptible to corrosion challenges.

Coating failure detection in scratch testing: A cross-sectional SEM/FIB microscopic study coupled with nonlinear analysis methods in a model titanium nitride/stainless steel system

In modern scratch testing, efforts are made to improve the quality of quantitative measurement of the coating adhesion. This paper proposes a new method for analyzing data collected in scratch tests. The nonlinear analysis methods of both penetration depth and tangential force time series were combined with SEM/FIB cross-sectional studies to monitor the progression of coating failure in a model titanium nitride coated stainless steel system. The applicability of recurrence quantification analysis and diagonal cross-recurrence analysis in scratch testing were discussed.

Oct4 promotes the progression and radioresistance of esophageal squamous cell carcinoma by regulating epithelial-mesenchymal transition

Objective: To explore the specific role and molecular mechanism of octamer-binding transcription factor 4 (Oct4) in promoting the progression of esophageal squamous cell carcinoma and radioresistance. Methods: The Gene Expression Profile Data Dynamic Analysis (GEPIA) database was used to analyze the expression differences of the Oct4 gene in different types of tumor tissues and their corresponding adjacent normal tissues. The clinical data and surgical resection tissue specimens of 196 patients with esophageal squamous cell carcinoma who received surgery combined with radiotherapy at Henan Provincial Chest Hospital from January 2013 to May 2022 were collected. Immunohistochemistry was used to detect the expression of Oct4 protein in the tumor and adjacent tissues. The lentiviral packaging system was used to construct esophageal squamous cell carcinoma cell lines that up-regulated or down-regulated Oct4. The cell counting kit 8 (CCK-8) was used to detect the cell proliferation ability, the scratch test was used to detect the cell migration ability, and the clone formation test was used to detect the cell radiosensitivity. Immunofluorescence experiment was used to detect DNA damage level, and Western blot was used to detect the expressions of Oct4, human phosphorylated histone (γ-H2AX), E-cadherin, N-cadherin, vimentin, and zinc finger E box binding homology box 1 (ZEB1). Results: The analysis of GEPIA database showed that the expression level of Oct4 mRNA in esophageal carcinoma was higher than that in paracancerous tissues. The expression level of Oct4 protein in tumor tissues was 78.35±1.42, which was higher than that in adjacent tissues (16.27±0.49). The survival time of patients with a high expression of Oct4 was significantly shorter than that of patients with a low expression of Oct4 (25.40 and 47.00 months). Compared with the control group, the proliferation ability of KYSE510 cells in the Oct4 up-regulated group was enhanced after 72-h culture, and the cell migration ability of these cells was also enhanced, with the migration rate being (41.67±1.20)% vs (23.67±1.86)% after 24-h culture. The radiosensitivity of cells in this group decreased, with the radiosensitivity enhancement ratio being 0.69±0.06 vs 1.00±0.02. After radiotherapy, the expressions of γ-H2AX and E-cadherin decreased, while the expressions of ZEB1, vimentin and N-cadherin increased. Compared with the control group, the proliferation ability of KYSE150 cells in the Oct4 down-regulated groups 1 and 2 decreased (absorbance being 2.51±0.17, 2.38±0.16, and 3.33±0.07, respectively, P＜0.01) after 72-h culture, and the migration ability also decreased, with the migration rate being (13.33±0.88)%, (13.00±1.00)%, and (40.33±2.03)%, respectively (all P＜0.001), after 24-h culture. The radiosensitivity was enhanced, with the radiosensitivity enhancement ratio being 1.34±0.11,1.24±0.07, and 1.00±0.02, respectively (all P＜0.05). After radiotherapy, the expressions of γ-H2AX and E-cadherin increased, while the expressions of ZEB1, vimentin and N-cadherin decreased. Compared with the control group, the proliferation ability of KYSE510 cells in the ZEB1 down-regulated group decreased [absorbance being 1.33±0.15 vs 1.81±0.16 (P=0.002)] after 72-h culture. The radiosensitivity was enhanced, with the radiosensitivity enhancement ratio being 1.37±0.11 vs 1.00±0.01 (P=0.037), and after radiotherapy the expression of γ-H2AX increased. Conclusion: Oct4 is involved in the regulation of epithelial-mesenchymal transformation of esophageal squamous cell carcinoma, which promotes the proliferation, migration, and radioresistance of esophageal squamous cell carcinoma.

Inhibition of miR-20a-5p Suppresses Epithelial-Mesenchymal Transition of Colorectal Cancer Cells Through GJA1.

This study was designed to clarify the role of GJA1 in colorectal cancer. qPCR was adopted to detect the GJA1 and miR-20a-5p expression levels in tumor tissues and cells; and EdU, Transwell assay, Scratch test to examine the migration, invasion, and proliferation of colorectal cancer cells. The EMT-related protein expression was measured by immunofluorescence and western Blot. The binding relationship between GJA1 and miR-20a-5p was examined using dual luciferase reporting subsystem. In situ hybridization was utilized to examine the miR-20a-5p expression in tumor tissues and metastases. Rescue experiments were performed by simultaneous transfection of sh-GJA1 inhibitor and miR-20a-5p inhibitor. The miR-20a-5p expression was high and the GJA1 expression was low in colorectal cancer tissues and cells. A targeting relationship was found in GJA1 and miR-20a-5p targets. The invasion, migration, and proliferation of colorectal cancer cells can be inhibited by overexpression of GJA1. Meanwhile, overexpression of GJA1 markedly elevated the e-cadherin expression, but reduced the levels of vimentin, α-SMA and n-cadherin expression. miR-20a-5p inhibitor + sh-GJA1 promoted the invasion, migration, and proliferation of colon cancer cells and EMT process. Overall, miR-20a-5p could target GJA1 to down-regulate the GJA1 expression, thereby regulating the EMT response, and ultimately promoting the progression of colorectal cancer.

Transcriptomics of tissue exosomes to investigate miR-195-5p’s amelioration of endometrial fibrosis via the YAP-Smad7 pathway: an animal study

BackgroundA significant research gap exists regarding the role of tissue exosomes in intrauterine adhesions (IUAs). This study aims to investigate the involvement of miR-195-5p and its regulatory network in IUAs through the analysis of tissue exosomes.MethodsExosomes from rat uterine tissue with intrauterine adhesions were analyzed via transcriptomics to identify downstream target genes of miR-195-5p, cross-referencing with the human endometrial transcriptomics database GSE224093. Dual luciferase labeling confirmed miRNA-target gene interactions. The therapeutic efficacy of a miR-195-5p agonist was assessed in vivo through HE staining, Masson staining, and mating tests. The mechanisms underlying extracellular matrix (ECM) deposition and myofibroblast transdifferentiation in endometrial fibrosis were investigated both in vitro and in vivo using RT-PCR, Western Blot, immunofluorescence, and immunohistochemistry. Migration ability of endometrial stromal cells was evaluated using CCK8, scratch tests, and Transwell assays. Finally, the clinical potential of miR-195-5p was compared with autologous adipose-derived mesenchymal stem cells.ResultsThe expression of miR-195-5p in uterine tissue exosomes from intrauterine adhesions was found to be decreased. Treatment with a miR-195-5p agonist resulted in improved endometrial health, reduced fibrosis, increased glandular density, and enhanced birth rates in rats. Both in vivo and in vitro experiments confirmed that miR-195-5p decreased ECM deposition, reduced myofibroblast transdifferentiation, and inhibited the migration of endometrial stromal cells. This was achieved through the downregulation of YAP expression in the Hippo pathway and the upregulation of Smad7. Notably, the therapeutic efficacy of miR-195-5p agonists was comparable to that of stem cell therapy, offering promising avenues for clinical application.ConclusionsDifferential expression of miR-195-5p in tissue exosomes can reduce ECM deposition and myofibroblast transdifferentiation, improving endometrial fibrosis by regulating the YAP-Smad7 pathway in the Hippo signaling cascade.

Sphingosine kinase-1 regulates migration and invasion of gastric cancer cells via targeting the nuclear factor-κB signaling pathway

To investigate the role of sphingosine kinase-1 (SPHK1) in regulating migration and invasion of gastric cancer (GC) cells. TIMER2.0, GEPIA and HPA databases were used to investigate SPHK1 expression in GC, and its association with prognosis of the patients was analyzed using Kaplan-Meier Plotter database. In 40 clinical GC and adjacent tissue samples, SPHK1 and MKI67 expressions were detected with immunohistochemistry, Western blotting, and RT-qPCR. Gene enrichment pathway analysis was conducted to explore the biological functions of SPHK1. In HGC-27 and MGC-803 cells, the effects of lentivirus-mediated SPHK1 knockdown or overexpression on cell migration and invasion and expressions of key proteins in the nuclear factor-κB (NF-κB) signaling were evaluated using cell scratch test, Transwell assays and Western blotting. The changes in tumorigenic capacity of the transfected GC cells were evaluated in nude mice. SPHK1 was highly expressed in GC tissues in negative correlation with overall survival, overall survival after progression, and relapse-free survival of the patients (all P<0.001). In clinical GC samples, SPHK1 and MKI67 expressions showed a positive correlation (P= 0.00049) and were both significantly up-regulated (P<0.001). Gene enrichment pathway analysis suggested the involvement of SPHK1 in cell adhesion, migration, angiogenesis and the NF-κB pathway (P<0.05). In the cell experiment, SPHK1 knockdown significantly decreased while SPHK1 overexpression enhanced migration and invasion abilities of the GC cells. SPHK1 positively regulated the expressions of phosphorylated P65 (P-P65), VEGFA and IL-17, and blocking the NF-κB pathway by PDTC significantly lowered migration and invasion ability of the cells. In nude mice, the GC cells with SPHK1 knockdown resulted in significantly reduced tumor size and mass, while the SPHK1-overexpressing cells showed enhanced tumorigenicity. SPHK1 regulates migration and invasion of GC cells via the NF-κB signaling pathway and may serve as a potential diagnostic marker for GC progression.

Novel biocompatible magnetron-sputtered silver coating for enhanced antibacterial properties and osteogenesis in vitro

Peri-implant infection is a serious complication in orthopedic surgery. This study aimed to reduce the incidence of peri-implant infection by developing a durable and safe antibacterial silver coating. We compared the antibacterial properties and process controllability of various coating techniques to identify the most effective method for silver coating. We refined substrate treatment techniques and coating thicknesses through antibacterial and scratch tests. Scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) were used to analyze the coating’s morphology and composition. Micron-sized magnetron sputtering silver coating samples underwent in vitro antibacterial testing, cytotoxicity testing, silver ion release testing, and osteogenic testing using membrane contact culture, CCK-8 assay, inductively coupled plasma (ICP) detection, and alkaline phosphatase (ALP) activity/osteogenic gene PCR. Magnetron sputtering demonstrated superior antibacterial properties, uniformity, and process controllability compared to alternative techniques. The optimal adhesion strength was achieved with a 0.5 μm coating thickness and a 400 mesh sandpaper pretreatment process, without compromising antibacterial efficacy. The coating showed near-perfect antiseptic results in antibacterial and anti-biofilm tests. Fibroblasts cultured in silver ion precipitation medium exhibited growth rates of 89% on day 30 and 88% on day 90, compared to 95% in the control group. The osteogenic test indicated that the magnetron sputtering silver coating promotes osteogenesis effectively. Our study demonstrated that micron-sized magnetron sputtering silver coating has potential for clinical use to prevent peri-implant infections in the future.